Enzyme Discovery & Production Technology Webinar

A webinar was held on Enzyme Discovery & Production Technology on 21st August 2015. The area of enzyme discovery has advanced incessantly since the late 1990s, a synergy of ever efficient/powerful gene cloning techniques, the logarithmic pace a genome sequencing and powerful/accessible bioinformatics tools. This webinar explores the recent contribution of Prozomix in this fast paced and competitive area, with respect to proprietary technologies and HT-methodologies developed for:

(1) Discovery of novel and highly diverse enzymes for biocatalysis applications

(2) HT-characterisation of large biocatalysis enzyme panels

(3) Scaled production of biocatalysis enzymes

The application and further development of these technologies within the FP7 Project “BIOOX” is presented, along with the emerging importance of metagenomics in the area of industrial biotechnology.

To watch the webinar, click here>>

KYRBIO recently held their end of project webinar titled 'The Discovery, Development and Demonstration of Biocatalysis for use in the Industrial Synthesis of Chiral Chemicals'. The webinar gives a brief overview of the four year FP7 project Kyrobio. The KYROBIO project has used an SME-focused approach to address industrially identified needs for chiral synthesis using biocatalysis with partners that have the potential to exploit the results generated. Our overarching challenge was that multiple chiral centres form a significant feature in several chosen chemicals. The control of reaction stereochemistry was targeted for added value of the Kyrobio technology. This leads into challenges in molecular biology, enzymology and process engineering to name a few. All these areas are covered in the webinar and for more information please contact ed.jones@ctechinnovation.com

University of Manchester Results Published in International Chemistry Journals

The work carried out at the University of Manchester as part of EU project KYROBIO's consortium has been recently concluded and disseminated in an international chemistry journal. Over the past four years researchers in Manchester (Dr. Nicholas Weise and Prof. Nicholas Turner) have sought to expand the suite of ammonia lyase enzymes used for the synthesis of unnatural amino acids as valuable pharmaceutical and fine chemical precursors. Investigation of a reported enzyme, EncP, in a biocatalytic context revealed it to be a good starting template. Using structure-aided rational design the enzyme was engineered and the amenability of its regioselectivity to fine-tuning demonstrated. This work has been recently published in the Journal of the American Chemical Society:

Weise, N. J., Parmeggiani, F., Ahmed, S. T., & Turner, N. J. (2015). The Bacterial Ammonia Lyase EncP: A Tunable Biocatalyst for the Synthesis of Unnatural Amino Acids. Journal of the American Chemical Society, 137(40), 12977–12983. doi:10.1021/jacs.5b07326

The analytical methods and biochemical insight involved in the completion of this project has led to collaborative efforts with researchers from another industrial-academic consortium (BBSRC sLOLA between the University of Manchester and pharmaceutical company Glaxo Smith Kline). As such this year KYROBIO work in Manchester has also been acknowledged in the following publications in leading chemistry journals:

Ahmed, S. T., Parmeggiani, F., Weise, N. J., Flitsch, S. L., & Turner, N. J. (2015). Chemoenzymatic Synthesis of Optically Pure l - and d -Biarylalanines through Biocatalytic Asymmetric Amination and Palladium-Catalyzed Arylation. ACS Catalysis, 5(9), 5410–5413. doi:10.1021/acscatal.5b01132

Parmeggiani, F., Lovelock, S. L., Weise, N. J., Ahmed, S. T., & Turner, N. J. (2015). Synthesis of D- and L-Phenylalanine Derivatives by Phenylalanine Ammonia Lyases: A Multienzymatic Cascade Process. Angewandte Chemie (International Ed. in English), 54, 4608–4611. doi:10.1002/anie.201410670

Kyrobio Researchers Contribute to Industrial Biotechnology MOOC

KYROBIO researchers has been involved in the coordination and delivery of a Massive Open Online Course (MOOC) in Industrial Biotechnology as part of the University of Manchester’s research beacons initiative. The course, which is due to be released worldwide to thousands of participants during the next year, is being designed to introduce students from diverse backgrounds to the principles of industrial biotechnology, with a focus on enzyme technology and biocatalysis for the production of pharmaceutical compounds. Submodules of the MOOC are being delivered by both internal and external academic and industrial partners, including members of KYROBIO and other EU-funded projects in this field.

Communication to Young Researchers and The Public

Nicholas Weise was selected to present his work within the KYROBIO project at a young researchers’ meeting at the University of Stuttgart, DE. The event, New Reactions with Enzyme and Microorganisms, was attended by PhD students and early career postdoctoral research scientists from across Europe. Nicholas delivered a talk on the use of phenylalanine ammonia lyase and aminomutase enzymes for the production of unnatural amino acids, covering work done within the consortium as well as collaborative projects with other researchers on these enzymes. The KYROBIO project was also represented at public engagement events from local initiatives, such as the Manchester Science Festival and Manchester Institute of Biotechnology high school student open day, to international events, such as the iGEM Giant Jamboree, Boston US. In this way the work of KYROBIO researchers in the context of biotechnology was highlighted by raising awareness and encouraging student participation in the use of enzymes to produce industrially and biomedically relevant compounds.

University of Stuttgart Update

Natural squalene hopene cyclases (SHCs) catalyze a sophisticated polycyclization reaction cascade that is started from the symmetrical substrate squalene to the products hopene and hopanol. This cyclization reaction is one of the most complex reactions known in biochemistry and reveals the incomparable high selectivity that can be obtained by performing reactions in enzyme´s active sites. The team of Prof. Bernhard Hauer at the Institute of Technical Biochemistry at the Universitaet Stuttgart, Germany used functionalized acyclic terpene derivatives as substrates for SHCs which enable the syntheses of numerous non-natural carbopolycyclic and heteropolycyclic products in a stereoselective manner. Various nucleophiles attacking the carbocationic intermediates in the reaction enabled the stereoselective syntheses of a multitude of polycyclic products. Furthermore, they demonstrated that SHCs can perform Friedel–Crafts alkylations with aromatics as well as hydroamidations revealing the potential of these enzymes for chiral Brønsted acid catalysis. The researchers started to direct the evolution of the SHC from Alicyclobacillus acidocaldarius (AacSHC) to catalyze novel carbon-carbon bond forming reactions while releasing the protonation machinery from its natural polycyclization chemistry. This yielded enzyme variants catalysing different non-natural Brønsted acid-catalyzed reactions in a highly selective manner. The development of novel, non-natural enzyme function is still an unexplored area and its design is an immense challenge in biocatalysis. The diversity of Brønsted acid-catalyzed reactions addressed will allow to catalyze a plethora of chemical reactions having no natural counterparts and to gain a greater fundamental understanding in discovering new enzyme activities in the lab. The generation of further enzyme catalysts for Brønsted acid reactions will facilitate the synthesis of new product molecules of high added value.

Profile of Nick Weise, PhD Student, University of Manchester

Nick read in Molecular Biology at the University of Manchester with specialisation in evolutionary biology and structural bioinformatics, graduating with a B.Sc. (Hons) and a university Award for Outstanding Academic Achievement in June 2012. During his university career he held student research scholarships at the Institute of Cancer Research, London, The MRC Laboratory of Molecular Biology, Cambridge, and the Wellcome Trust Sanger Institute, Hinxton. Nick was recruited onto the KYROBIO project in July 2012, beginning doctoral studies at the Centre of Excellence in Biocatalysis. - part of the Manchester Institute of Biotechnology. His research is focussed on the engineering and implementation of ammonia lyase / aminomutase enzymes in the synthesis of unnatural amino acids. Nick is also a keen science communicator, participating in and organising various public engagement events both in Manchester and across the UK, as well as contributing to dissemination activities within the KYROBIO consortium.

Transam 2.0: Chiral Amines through (Bio)Catalysis - A thoughtpiece by Eva Fischereder, PhD Student, University of Graz

"I had the opportunity to give an oral presentation of my PhD-project this year at the Transam 2.0 in Greifswald. The topic of the conference fitted very nicely to my PhD project, since the conference focused on the synthesis of chiral amines through biocatalysis. The target amines I concentrated on in my talk were strictosidine derivatives which I prepared via a diastereoselective cascade reaction employing two enzymes namely a ?-transaminase and a strictosidine synthase. This conference gave me the opportunity to get used to present in front of a big audience, and allowed me to meet and communicate with people from industry and academia. Furthermore I got an inside into the state of the art scientific achievements in this area as well as several positive suggestions for my thesis. Since various members of the KYROBIO project also attended the conference we had the opportunity to scientifically exchange besides the fixed meeting dates. For me the Transam had an inspiring and fairly personally atmosphere and I am looking forward for the next one."

KYROBIO Project Presents at BIOCAT 2014

The 7th International Congress on biocatalysis (biocat2014) took place from August 31st to September 4th 2014 and was hosted by the Technical University of Hamburg, Germany.

400 attendees from 46 nations, both from academia and industry, presented and discussed current developments in biocatalysis. Over 50 lectures were given and almost 300 posters were presented. The conference gave a great opportunity to the people from the Kyrobio project to present their work. Kerstin Steiner was invited to give a lecture about the potential of enzymes with cupin fold in biocatalysis. Elisa Lanfranchi, Eva Fischereder, Hesam Arabnejad, Nick Weise and Jörg Dominicus presented posters about their work within the Kyrobio project.

During several poster sessions it was possible to discuss ones research with other scientists as well as to explore other research fields presented by others. The conference dinner took place on a ship and gave the opportunity to meet people and discuss with them in a relaxed atmosphere. Altogether the biocat2014 was a great event to learn about the latest topics in biocatalysis and to meet other scientists in the field of biocatalysis.

Project Partner 'University of Stuttgart' Produces WEBINAR for KYROBIO

The University of Stuttgart is situated in the middle of a highly dynamic economic region with a worldwide reputation for excellence in the fields of mobile and information technology, production, process engineering as well as in life sciences. Professor Bernhard Hauer, Institute of Technical Biochemistry, from the University of Stuttgart discusses "Squalene Hopene Cyclases" in this webinar.

To watch the webinar, click here>>

KYROBIO Results Disseminated at a High Profile Showcase in Brussels

Business leaders, policy makers and academics came together on December 3rd 2013 to hear about the leading developments in industrial biotechnology. The day was organised to disseminate the mid-stage results of the KYROBIO project and the conclusion of a related the three-year FP7 programme, BIONEXGEN. ‘The next generation of biocatalysis for industrial chemical synthesis’ and provided an opportunity for people to:

• Understand the technology developments from key EU funded projects
• Come together with the EU IB community
• See the impact of EU funding and the potential of IB
• Identify the value-add of a network of collaborative EU funded projects

Throughout the day the event brought to the forefront the broad and varied area of biocatalysis, from novel process routes to new functionality and applications. It highlighted the impact of FP7 funded collaborative research, with presentations from leading academics and successful businesses participating in these projects. Click on the links below to view presentations and further information:

European Community Discuss Future Direction for Colalboration in Industrial Biotechnology
Enzymatic Amine Synthesis - Prof. Nick Turner, University of Manchester
IB Enabling Technologies - Prof. John Woodley, Technical University of Denmark
Protein expression: key technologies for enzyme engineering and chemical production by IB - Prof. Anton Glieder, Austrian Centre for Industrial Biotechnology
Funding for IB in Horizon 2020 - Johanna Dupont-Inglis, EuropaBio

Published Paper: Deracemization By Simultaneous Bio-oxidative Kinetic Resolution and Stereoinversion

Prof. Dr. Wolfgang Kroutil, University of Graz (Kyrobio Project Partner) is one of the Authors of the above contribution. Abstract: Deracemization, that is, the transformation of a racemate into a single product enantiomer with theoretically 100 % conversion and 100 % ee, is an appealing but also challenging option for asymmetric synthesis. Herein a novel chemo-enzymatic deracemization concept by a cascade is described: the pathway involves two enantioselective oxidation steps and one non-stereoselective reduction step, enabling stereoinversion and a simultaneous kinetic resolution. The concept was exemplified for the transformation of rac-benzylisoquinolines to optically pure (S)-berbines. The racemic substrates were transformed to optically pure products (ee>97 %) with up to 98 % conversion and up to 88 % yield of isolated product.

Read more>>

Public Engagement

A team of EU-funded scientists from the University of Manchester have been engaging in outreach events to increase public understanding of enzymes and awareness of their use in the chemical and biotechnology industries. The team features researchers from biocatalysis-focused projects such as AMBIOCAS, SUPRABIO, BIONEXGEN and of course KYROBIO. The aim of these dissemination initiatives has been to highlight the importance of EU funding in this field to a wider audience by demonstrating how the research carried out can have a real impact in food processing, bioenergy and synthesis of pharmaceuticals and other fine chemicals. The enzyme outreach activities have so far been represented at larger public engagement festivals, such as Manchester Science Spectacular and additional events at the Museum of Science and Industry. Use of simple, hands-on enzyme experiments combined with more in depth reading material about enzymes in everyday life allows the outreach efforts to be catered to people of all ages and backgrounds.

Published Paper: New Opportunities by Synthetic Biology for Biopharmaceutical Production in Pichia Pastoris

  • Vogl Thomas, Hartner Franz S., Glieder Anton: New opportunities by synthetic biology for biopharmaceutical production in Pichia pastoris. (2013) Current Opinion in Biotechnology, 24, 1094-1101; DOI: 10.1016/j.copbio.2013.02.024

Biopharmaceuticals are an integral part of modern medicine and pharmacy. Both, the development and the biotechnological production of biopharmaceuticals are highly cost-intensive and require suitable expression systems. In this review we discuss established and emerging tools for reengineering the methylotrophic yeast Pichia pastoris for biopharmaceutical production. Recent advancements of this industrial expression system through synthetic biology include synthetic promoters to avoid methanol induction and to fine-tune protein production. New platform strains and molecular cloning tools as well as in vivo glycoengineering to produce humanized glycoforms have made P. pastoris an important host for biopharmaceutical production.

Read more>>

Presentations of KYROBIO Results by ACIB (Project Partner)

Title: “Engineering of cupin hydroxynitrile lyases” oral presentation by Romana Wiedner, Bettina Kothbauer, Mandana Gruber-Khadjawi, Helmut Schwab, Kerstin Steiner  at ECB 16, 2014 July 13th - 16th in Edinburgh, Scotland

Title: “Blue Native PAGE: bioprospecting for new hydroxynitrile lyases” presentation by Elisa Lanfranchi, Eva M. Köhler, Kerstin Steiner, Anton Glieder , Margit Winkler at biocat 2014, 2014 Aug. 31st – Sept. 4th in Hamburg, Germany